Kit for Chinese pediatric asthma susceptibility gene SNP (single nucleotide polymorphism) genotyping and application method of kit for Chinese pediatric asthma susceptibility gene SNP genotyping

A technology for susceptibility genes and childhood asthma, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as application limitations, low accuracy, and cumbersome operations.

Inactive Publication Date: 2016-01-13
LIUZHOU CITY HEALTHCARE HOSPITAL FOR WOMEN & CHILDREN
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Problems solved by technology

However, this method has disadvantages such as time-consuming, cumbersome operation, and low accuracy.
Some scholars also use real-time fluorescent quantitative PCR combined with DNA sequencing methods to detect gene polymorphism sites, but this method is limited in the application of large-scale population screening or detection of multiple genes and multiple sites

Method used

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  • Kit for Chinese pediatric asthma susceptibility gene SNP (single nucleotide polymorphism) genotyping and application method of kit for Chinese pediatric asthma susceptibility gene SNP genotyping
  • Kit for Chinese pediatric asthma susceptibility gene SNP (single nucleotide polymorphism) genotyping and application method of kit for Chinese pediatric asthma susceptibility gene SNP genotyping
  • Kit for Chinese pediatric asthma susceptibility gene SNP (single nucleotide polymorphism) genotyping and application method of kit for Chinese pediatric asthma susceptibility gene SNP genotyping

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Effect test

Embodiment 1

[0077] Example 1: As shown in the accompanying drawing, the monitoring of SNP typing of susceptibility genes in asthmatic patients:

[0078] Sample selection: The samples were obtained from the DNA library of asthmatic patients in the Department of Internal Medicine of Liuzhou Maternal and Child Health Hospital. Among them, there were 100 males and 100 females. All patients in the bank signed the informed consent before the samples were collected, and promised to cooperate with the follow-up diagnosis and follow-up.

[0079] 1. PCR amplification of 9 segments of 8 asthma susceptibility genes with a single pair of primers

[0080] The PCR reaction system is 20 μL, including 10×PCR buffer (Mg 2+ free) 2 μL, 0.8 μL MgCl 2 (50mM), dNTP mix 0.5μL (10mM), 0.2μL PlatinumTaq DNA polymerase (5U), forward primer 0.5μL, reverse primer 0.5μL, 1μL DNA template (20mg / L), 14.5μL ddH 2 O. The PCR reaction conditions were: denaturation at 95°C for 2 min, denaturation at 95°C for 30 s, anne...

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Abstract

The invention relates to a kit for Chinese pediatric asthma susceptibility gene SNP (single nucleotide polymorphism) genotyping and an application method of the kit for Chinese pediatric asthma susceptibility gene SNP genotyping, in particular to a kit for simultaneous qualitative genotyping of nine SNP loci of eight Chinese pediatric asthma susceptibility genes and an application method of the kit for simultaneous qualitative genotyping of the nine SNP loci of the eight Chinese pediatric asthma susceptibility gene. Genotypes of the nine SNP loci are obtained simultaneously by taking an rs1042713 locus of an ADRB2 gene, an rs1805010 locus of an IL-4R alpha gene, an rs20541 locus of an IL-13 gene, an rs1800925 locus of the IL-13 gene, an rs2495636 locus of an IL-13R alpha gene, an rs2243250 locus of an IL-4 gene, an rs1800629 locus of a TNF-alpha gene, an rs569108 locus of an MS4A2 gene and an rs528557 locus of an ADAM33 gene as detection objects, designing amplification primers and extension primers respectively aiming at mutation of the nine SNP loci, performing multiplex PCR (polymerase chain reaction) amplification and marker extension aiming at the nine SNP loci, and performing capillary electrophoresis. The kit for SNP genotyping is convenient to use, simple to operate, low in cost, high in flux and direct and reliable in detection results and is applicable to large-scale screening of Chinese pediatric asthma susceptibility gene SNP genotyping.

Description

technical field [0001] The invention relates to the field of SNP typing detection in the technical field of genetic engineering, in particular to a Chinese children's asthma susceptibility gene SNP typing kit and a use method thereof. Background technique [0002] Asthma is a common chronic respiratory disease that seriously endangers children's health. It usually occurs in winter and spring, and the first onset is mostly between 0-5 years old. Its morbidity rate is high, and it often manifests as a recurrent chronic course of disease, which seriously affects the study, life and activities of children, affects the growth and development of children and adolescents, and also brings a heavy burden to the families of children. Due to untimely or inappropriate treatment, many children with asthma develop into adult asthma with protracted recovery, impaired lung function, and some patients even completely lose their ability to perform physical activities. Severe asthma attacks c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q2531/113C12Q2537/143C12Q2521/319C12Q2521/525C12Q2533/101
Inventor 严提珍龙易勤曾定元唐宁王麟郑敏李伍高李哲涛余宾罗世强廖燕玲
Owner LIUZHOU CITY HEALTHCARE HOSPITAL FOR WOMEN & CHILDREN
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